Objective To investigate the phenotype, frequency of Th17 cells and the association between Th17 cells and viral clearance in patients with H1N1 influenza A. Methods Three groups including 70 confirmed patients with H1N1 influenza A, 30 patients with seasonal influenza as well as 68 healthy subjects as controls were enrolled in this study. The percentages of Th1, Th2, Treg and Th17 lymphocytes in the peripheral blood were determined by intracellular staining and flow cytometry. The levels of interferon-γ (IFN-γ), transforming growth factor-beta (TGF-β),interleukin-6 (IL-6) in plasma and supernatant of the peripheral blood mononuclear cell (PBMC)culture were quantified by enzyme-linked immunosorbent assay (ELISA). Viral load in nasopharyngeal swabs was detected by real time quantitative reverse transcription-polymerase chain reaction (RTPCR). Data were analyzed by one way ANOVA and liner correlation analysis. Results The percentage of Th17 cells in H1N1 influenza A patients was (2. 740±0. 210)%, which the percentage of was significantly decreased compared to healthy subjects (3. 443 ±0. 154)% and seasonal influenza patients (3. 443±0. 277) % (F=4. 242, P＜0. 05); while the percentage of Thl, Th2 and Treg cells were not significantly different among these groups. Moreover, the TGF-β level in plasma of H1N1 influenza A patients was (10±8) ng/mL, which was significantly lower than healthy subjects (43 ±32 ) ng/mL and seasonal influenza patient ( 18 ± 10) ng/mL ( F= 17.72, P＜0.01 ). The TGF-β level in the supernatant of PBMC culture of H1N1 influenza A patients was (782 ± 736) pg/mL, which was significantly lower than healthy subjects (1462±315) pg/mL and seasonal influenza patients (1481 ±348) pg/mL (F=5. 730, P＜0.01). Additionally, the viral clearance period was inversely correlated with the percentage of Th17 cells (r=-0.38, P=0.02). Conclusions The proportion of Th17 cells in patients with H1N1 influenza A is significantly decreased, which is closely correlated with the level of TGF-β. This decrease may results in the delayed viral clearance.

Objective To Screen for safe and effective vaccine candidates for EV71,provide a theoretical basis for development of EV71 vaccines in the future.Methods VP1 gene of enterovirus was used to design a target for development of EV71 vaccines.Different vaccine candidates,including inactivated EV71 vaccines,VP1 protein vaccine,DNA vaccines of different doses,were used to challenge female BALB/c mice by intramuscular injection at baseline (0),2 weeks,4 weeks,and caudal vein blood was collected at 0,2,4,6,8,10,and 16 weeks,and BALB/c mice were sacrificed and the spleen cells were collected for detection of both humoral immunity and cellular immunity to evaluate the efficacy and safety of the vaccine candidates.Results IgG antibody titers were increased at 2 weeks,remarkably increased at 4 weeks,reached a peak at 8 weeks,at least sustained for 16 weeks during the whole observation period,subtypes of IgG1 and IgG2a were the major component.The three vaccines could induce cellular immunity characterized by EV71 specific γ-IFN and IL-4 production.Our results indicated that inactivated EV71 vaccine was superior to the other vaccine candidates.Conclusions Inactivated EV71 vaccines,VP1 protein vaccine,DNA vaccines can induce both strong and sustainable humoral and cellular immunities in challenged mice,and the inactivated EV71 vaccine is superior to the other vaccine candidates,which needs to be proved their immunity by challenge assay in the future.

Objective To analyze clinical and laboratory features, viral load and viral shedding period of patients with mild or severe H1N1 influenza A infection. Methods Seventy mild cases and 16 severe cases with concurrent pneumonia were included from Shcnzhen area for analysis.Nasopharyngeal-swab specimens of patients were collected and viral load was detected by real-time quantitative polymerase chain reaction (PCR) assay during their hospitalization. The viral load and viral shedding period were compared between patients over 14 years old and less than 14 years old, and between 70 mild cases without pneumonia and 16 severe cases with pneumonia. The statistic analysis was performed using t test and chi square test. Results The most common symptoms and signs of the patients were fever, cough and enlargement of tonsils. However, the severe cases suffered more frequently from cough, dyspnea and high fever compared with the mild cases (x2 = 10. 9 and 14.3, respectively, t=3.65; both P＜0.01 ). The levels of white blood cell (WBC) count and alanine arninotransferase (ALT) of severe patients were both significantly higher than those of mild patients(t= 3.2, 2.4,respectively; both P＜0.05). The chest radiology of the severe cases showed interstitial pneumonia,mostly with ground glass image. The viral load of patients under 14 years was significantly higher than those over 14 years [(4.86± 1.23) lg vs (4. 17±0.89) lg; t=2.3, P＜0.05], and the viral shedding period of patients under 14 years was significantly longer than those over 14 years [(5.33±0. 49) d vs(3. 63±0.28) d; t=3.4, P＜0.01]. The severe patients also displayed significantly higher viral load and prolonged viral shedding period than the mild patients [(6. 36±1. 44) lg vs (4. 35±0.99) lg, t=6.1,P＜0.01; (5.75±1.77) d vs (4. 24±1. 96) d, t=3.2, P＜0.01]. Conclusion Age anddisease severity of patients with H1N1 influenza A infection are significantly associated with viral load and viral shedding period.

Objective:To understand the diagnostic value of tuberculosis (TB) pathogenic detection methods (TPDM) in pathological tissue for TB.Methods:A retrospective study was conducted with 190 pathological specimens from different tissues suspected with TB from Third People′s Hospital of Shenzhen during May 2016 and May 2019. Specimens were divided into four groups according to histomorphology: group one, necrotizing granulomatous inflammation (109 cases); group two, non-necrotic granulomatous inflammation (20 cases); group three, non-granulomatous inflammation (45 cases); group four, non-tuberculous lesions (16 cases). The positive rates of each TPDM among specimens from four groups were compared. The positive rates of all TPDM for specimens from group one were compared. Meanwhile, the influence of antituberculosis treatment course on the TPDM was analyzed. Chi-square test or Fisher′s exact test was used for statistical analysis.Results:The positive rates of Ziehl-Neelsen acid-fast staining among the four groups were 17.4%(19/109), 5.0%(1/20), 4.4%(2/45) and 0(0/16), respectively. The positive rates of Mycobacterium tuberculosis (MTB) complex culture were 32.0%(32/100), 4/19, 4.8%(2/42) and 0(0/16), respectively. The positive rates of Mycobacterium tuberculosis/rifampin resistance real-time quantitative nucleic acid amplification detection system (Xpert MTB/RIF) were 74.3%(81/109), 15.0%(3/20), 13.3%(6/45) and 0(0/16), respectively. The positive rates of fluorescent quantitative polymerase chain reaction (FQ-PCR) were 63.0%(58/92), 0(0/15), 2.6%(1/38) and 0(0/10), respectively. The positive rates of simultaneous amplification and testing (SAT) were 32.4%(24/74), 0(0/10), 0(0/15) and 0(0/10), respectively. The differences of each TPDM among four groups were all statistically significant (all P<0.05). The positive rate of Xpert MTB/RIF in group one specimens was significantly higher than those of acid-fast staining, MTB culture and SAT ( χ2=71.016, 37.162 and 35.679, respectively, all P<0.01), while the difference was not statistically significant when compared with FQ-PCR ( χ2=2.517, P=0.112). The positive rate of combined TPDM (85.3%(93/109)) was significantly higher than Xpert MTB/RIF(74.3%(81/109)) ( χ2=4.100, P=0.043). The positive rates of acid-fast staining group 1A (anti-tuberculosis treatment course was less than one month) and group 1B (anti-tuberculosis treatment course was longer than one month) were 14.3%(7/49) and 20.0% (12/60), respectively ( χ2=0.612, P=0.434); those of MTB culture were 48.9% (22/45) and 18.2% (10/55), respectively ( χ2=10.721, P=0.001); those of Xpert MTB/RIF were 69.4%(34/49) and 78.3%(47/60), respectively ( χ2=1.131, P=0.287); those of FQ-PCR were 55.0%(22/40) and 69.2%(36/52), respectively ( χ2=1.965, P=0.161); those of SAT were 43.3%(13/30) and 25.0%(11/44), respectively ( χ2=2.736, P=0.098). Conclusions:The results of TPDM correlate closely with the typical histomorphological features of tuberculosis. Xpert MTB/RIF possesses significantly higher sensitivity than any other single TPDM, and is not attenuated by early anti-tuberculosis treatment. Combined TPDM could significantly improve the sensitivity of TB pathogenic detection, which is suggested to be applied when the tissue specimen is sufficient.

Objective To explore the role of Entecavir in CD4 * T cells and CD4 + CD25 + Foxp3 + regulatory T cells of patients with chronic hepatitis B.Methods Patients with chronic active hepatitis B (CAH,n =50) were divided into Entecavir treatment group (n =30) and controls (n =20).The entecavir treatment group and the controls were followed up for 12 months.The frequency and phenotype of peripheral CD4 + and CD4 + CD25 + Foxp3 + T cells were detected by flow cytometry.Results Patients with CHB receiving entecavir treatment for 3 months,6 months and 12months exhibited significantly higher frequency of CD4 + T cells and CD4/CD8 ratio as opposed to those of baseline levels(P ＜0.05).The frequency of CD4 +CD25 + Foxp3 + T cells from the patients receiving entecavir treatment for 3 months,6 months and 12 months was significantly decreased compared with that of basline level(P ＜ 0.05).However,the frequency of CD4 T cells and CD4 + CD25 + Foxp3 + T cells,as well as CD4/CD8 ratio of the control group following up for 12 months displayed no significant differences compared with its baseline levels.The serum HBV DNA level from the baseline to Entecavir treatment for 12 months was correlated with the frequency of CD4 + CD25 +Foxp3 * T cells from the baseline to Entecavir treatment for 12 months.Conclusions Entecavir not only can effectively suppress HBV replication,but also simultaneously improve the cellular immune function of chronic hepatitis B.

Objective:To evaluate the efficacy of daily use of a test emollient combined with topical glucocorticoids applied at the weekend for delaying the recurrence of atopic dermatitis (AD) in children during the maintenance period.Methods:A randomized, blank-controlled, multicenter clinical study was conducted in children with moderate AD from Beijing Children′s Hospital, Capital Medical University, Children′s Hospital of Chongqing Medical University and Shenzhen Children′s Hospital from March 2021 to February 2022. A total of 127 children aged 0 - 12 years with moderate AD were treated with topical glucocorticoids combined with emollients during the run-in period, 112 out of them achieved the investigator′s global assessment (IGA) score ≤ 1 point, and then the 112 patients were randomly divided into a test group (56 cases) and a control group (56 cases) at a ratio of 1∶1. Patients in the test group received treatment with a test emollient twice a day in combination with topical glucocorticoids applied at the weekend, and those in the control group were only treated with topical glucocorticoids at the weekend. Patients in the two groups were followed up at baseline, week 2 (± 3 d), week 4 (± 5 d), and week 12 (±7 d), as well as at the time of AD relapse, and the effect of the test emollient on the remission rate of AD in children during the maintenance period was evaluated, so were its effects on the dosage of topical glucocorticoids, pruritus, sleep, and skin pH. The occurrence of treatment-related adverse events was evaluated and recorded at the same time. Study endpoints were defined as AD relapse during the maintenance period, end of 12-week follow-up, or occurrence of serious adverse events. Comparisons of efficacy indicators between groups were conducted by using chi-square test, Kaplan-Meier survival analysis, Satterthwaite t′ test and Mann-Whitney U test. Results:In the full-analysis set, 45 (80.36%) patients with AD maintained remission in the test group (56 cases) and 30 (53.57%) in the control group (56 cases), and the remission rate difference between the two groups was 26.79% (95% confidence interval [ CI]: 10.09%, 43.49%; χ2 = 9.11, P = 0.003) ; the 12-week follow-up during the maintenance period showed that the time to first relapse was 75.05 ± 25.07 days in the test group, which was significantly longer than that in the control group (49.55 ± 33.92 days, t′ = 4.52, P < 0.001). At the study endpoint, the test group showed significantly decreased AD disease severity score (eczema area and severity index [EASI] score: 0.00 [0.00, 1.20] points vs. 0.60 [0.00, 4.00] points), pruritus visual analog scale (VAS) score (0.00 [0.00, 2.00] points vs. 2. 00 [0.00, 10.00] points), and sleep VAS score (0.00 [0.00, 0.00] points vs. 1.00 [0.00, 4.00] points) compared with the control group ( Z = -2.77, 2.43, 3.48, P = 0.006, = 0.015, < 0.001, respectively), while there was no significant difference in the pH value at the lesional sites between the test group and control group ( t = 0.97, P = 0.335). For the group aged 0 - 2 years, the average daily glucocorticoid dosage at the weekend in AD children during the maintenance period was significantly lower in the test group than in the control group ( Z = -1.97, P = 0.049) ; for the group aged >2 - 12 years, there was no significant difference in the average daily glucocorticoid dosage at the weekend between the two groups ( Z = -0.25, P = 0.802). During the study period, no significant difference was observed in the incidence of treatment-related adverse events between the test group (2/56, 3.57%) and control group (3/56, 5.36%; P = 1.000), and no serious adverse events occurred. Conclusion:Compared with the weekend treatment with topical glucocorticoids alone, the daily use of the test emollient combined with topical glucocorticoids at the weekend could markedly improve the remission rate of AD, prolong the time to relapse, and reduce the disease severity at relapse in children with AD during the maintenance period, which provides a new option for maintenance treatment of children with AD.